Western blot analysis of extracts from MOLT-4 and C2C12 cells, untreated (-) or treated with the PLK4 inhibitor CFI-400945 (500 nM, 18 hr; +), using PLK4 (E6A7R) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
PLK4 (E6A7R) Rabbit mAb recognizes endogenous levels of total PLK4 protein. An unknown band at 170 kDa has been observed.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu678 of human PLK4 protein.
At least four distinct polo-like kinases exist in mammalian cells: PLK1, PLK2, PLK3, and PLK4/SAK (1). PLK4/SAK is transcriptionally repressed by p53 and may contribute to p53-mediated apoptosis (2). PLK4 has also been identified as a key regulator of centriole duplication (3-5). PLK4 is typically expressed at low levels and is controlled by autoregulated instability such that treatment with PLK4 inhibitors, such as CFI-400945, leads to an increase in PLK4 expression (6,7). PLK4 is aberrantly expressed in multiple cancers and has emerged as a potential prognostic and therapeutic target (8).
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